RP-HPLC法同时测定熟地黄中5个核苷类成分的含量

目的: 建立同时测定熟地黄中次黄嘌呤、尿苷、腺嘌呤、鸟苷和腺苷5个核苷类成分的RP-HPLC法。方法: 采用Diamonsil C₁₈(4.6 mm×250 mm,5 μm)色谱柱,以乙腈-0.04 mol·L^-1磷酸二氢钾溶液为流动相梯度洗脱,流速1.0 mL·min^-1,检测波长254 nm;柱温30 ℃。结果: 次黄嘌呤、尿苷、腺嘌呤、鸟苷和腺苷浓度分别在1.0～16.0 μg·mL^-1(r=0.9999)、5.0～80.0 μg·mL^-1(r=0.9999)、1.0～16.0 μg·mL^-1(r=0.9997)、1.25～20.0 μg·mL^-1(r=0.9999)、1.0～16.0 μg·mL^-1(r=0.9999)范围内线性良好。加样回收率为98.1%～101.0%。结论: 该方法可以用于熟地黄中次黄嘌呤、尿苷、腺嘌呤、鸟苷和腺苷的含量测定。

Objective: To establish a RP-HPLC method for simultaneously determining five nucleosides (hypoxanthine, uridine, adenine, guanosine and adenosine) in Radix Rehmanniae Preparata. Methods: The Diamonsil C₁₈ (4.6 mm×250 mm, 5 μm) column was adopted and diluted with acetonitrile-0.04 mol·L^-1 potassium dihydrogen phosphate gradiently at the wavelength of 254 nm.The flow rate was 1.0 mL·min^-1 and the column temperature was 30 ℃. Results: The method was developed with a good linearity within the range of 1.0-16.0 μg·mL^-1 (r=0.9999), 5.0-80.0 μg·mL^-1 (r=0.9999), 1.0-16.0 μg·mL^-1 (r=0.9997), 1.25-20.0 μg·mL^-1 (r=0.9999) and 1.0-16.0 μg·mL^-1 (r=0.9999) for hypoxanthine, uridine, adenine, guanosine and adenosine, respectively.And the average recoveries of the five nucleosides were 98.1%-101.0%. Conclusion: The established method can be used for the quantification of five nucleosides(hypoxanthine, uridine, adenine, guanosine and adenosine)in Radix Rehmanniae Preparata.